scholarly journals Evaluation of Flow Cytometry and Kleihauer Techniques for Quantification of Fetomaternal Hemorrhage: A Prospective Cohort Study in Southwestern Iran

2018 ◽  
Vol 3 (4) ◽  
pp. 118-122
Author(s):  
Zeinab Keshavarz ◽  
Leili Moezzi ◽  
Reza Ranjbaran ◽  
Abbas Behzad-Behbahani ◽  
Masooma Abdullahi ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Prospective Cohort ◽  
Maternal Blood ◽  
Clinical Samples ◽  
Direct Immunofluorescence ◽  
Maternal Circulation ◽  
Cohort Design ◽  
Fetomaternal Hemorrhage ◽  
Negative Woman ◽  
Southwestern Iran

Background: Quantification of fetal red blood cells (RBCs) in maternal blood is of great importance to calculate appropriate dose of post-deliver anti D immunoglobulin in a rhesus D (RhD)-negative woman. Objective: The aim of this study is to evaluate a direct immunofluorescence flow cytometry technique in artificial and clinical samples and compared it to the Kleihauer-Betke test (KBT). Methods: This study was a prospective cohort design. Blood samples from 26 pregnant women who gave birth to RhD positive babies were tested using direct immunofluorescence flow cytometry and KBT techniques to determine the amount of FMH in the maternal circulation. The zone of D-positive cells was identified employing artificial samples including 0.3%, 0.6%, 1%, 1.5%, 2%, 5%, 10%, and 50% of D-positive fetal cells in D-negative maternal cells. Results: Analysis of 26 clinical samples for FMH showed consistent quantification with the flow cytometry and Kleihauer techniques. Although a good correlation was found between the KBT and flow cytometry results, in artificial samples containing more than 2% of fetal RhD positive cells, the flow cytometry results were closer to theoretical percentages. In a patient with FMH >4 mL, the FMH and consequently the required vial of Ig were overestimated using KBT. Conclusion: Most of the FMH calculated could have been neutralized by doses less than 625 IU, whereas the routine dose in Iran is more than double that amount (1500 IU). This achievement demonstrates that adjusting between the RhD immune globulin (RhDIg) dose and FMH size is inevitable.

Placental Pathologic Features in Fetomaternal Hemorrhage Detected By Flow Cytometry

2017 ◽  
Vol 20 (2) ◽  
pp. 142-151 ◽  
Author(s):  
Natasha E Lewis ◽  
Laura Marszalek ◽  
Linda M Ernst
Keyword(s):  
Flow Cytometry ◽  
Vascular Pathology ◽  
Clinical Effects ◽  
Placental Pathology ◽  
Fetal Blood ◽  
Maternal Circulation ◽  
Flow Cytometric ◽  
Fetomaternal Hemorrhage ◽  
Pathologic Features ◽  
Fetal Vessels

Background Fetomaternal hemorrhage (FMH) is a poorly understood entity that can have significant clinical effects. Flow cytometry is a reliable and relatively new method for FMH diagnosis. The objective of this study was to correlate placental pathology with FMH detected by flow cytometry. Methods All patients with available placentas and FMH flow cytometric testing performed from 2009 to 2015 were retrospectively reviewed. Cases were defined as ≥0.10% fetal red blood cells (RBCs) in the maternal circulation while controls contained <0.10%. Placental findings associated with FMH were determined. Results In this study, 35 cases and 79 controls were identified. Villous dysmaturity/immaturity was significantly more prevalent among the cases compared to the controls. Placentas with villous edema and nucleated RBCs (nRBCs) in fetal vessels were associated with greater mean volumes of fetal blood in the maternal circulation. Fetal and maternal vascular pathology was more frequent in the controls. When the cases were stratified into mild (<30 mL), moderate (30 mL–100 mL), and severe (>100 mL) FMH, nRBCs, villous dysmaturity/immaturity, and villous edema were all positively correlated with increasing FMH severity. The cases were more likely than the controls to display ≥2 of these 3 features. Fetal nRBCs within fetal vessels were semi-quantified and moderate to severe numbers of nRBCs were associated with higher mean volumes of fetal blood in maternal circulation. Conclusions Villous dysmaturity/immaturity, villous edema, and nRBCs in fetal vessels, findings compatible with fetal anemia, in addition to relatively few chronic placental changes, are the most significant placental findings in FMH detected by flow cytometry.

Quantification of fetomaternal hemorrhage: a comparative study of the manual and automated microscopic Kleihauer-Betke tests and flow cytometry in clinical samples

2004 ◽  
Vol 191 (2) ◽  
pp. 551-557 ◽  
Author(s):  
Denise M. Pelikan ◽  
Sicco A. Scherjon ◽  
Wilma E. Mesker ◽  
Godelieve M. de Groot-Swings ◽  
Geeske G. Brouwer-Mandema ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Comparative Study ◽  
Clinical Samples ◽  
Fetomaternal Hemorrhage

Approaching stability challenges for flow cytometry in a regulated bioanalytical environment

Bioanalysis ◽  
2019 ◽  
Vol 11 (20) ◽  
pp. 1845-1858 ◽  
Author(s):  
Anamica Muruganandham ◽  
Carolyne Dumont ◽  
Kuiyi Xing
Keyword(s):  
Flow Cytometry ◽  
Critical Parameter ◽  
Storage Period ◽  
Reliable Data ◽  
Clinical Samples ◽  
Sample Collection ◽  
Testing Laboratory ◽  
Stability Parameters ◽  
Stability Issues ◽  
Limited Period

Stability of samples for flow cytometry is a critical parameter since storage period of samples is restricted to only a limited period after collection. For most studies, clinical samples have to be shipped to a testing laboratory, in contrast to preclinical samples, which can be analyzed on-site or off-site. Therefore, evaluating stability is critical to provide flexibility on testing of samples to obtain reliable data. A wide variety of factors contributes to establishing stability from sample collection through acquisition. We provided suggestions for experimental and stability parameters to be taken into consideration when designing a flow cytometry method. The case studies presented represent how certain stability issues were overcome to perform flow cytometry assays in a regulated bioanalytical environment.

Flow cytometry inmunophenotyping can identify and characterize solid tumour cells in clinical samples: A preliminary study

2019 ◽  
Vol 493 ◽  
pp. S148-S149
Author(s):  
C. Quirós ◽  
L. Fernández ◽  
H. Torres ◽  
F. Domínguez ◽  
F.V. Álvarez ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Solid Tumour ◽  
Tumour Cells ◽  
Clinical Samples ◽  
Preliminary Study

scholarly journals Artifacts associated with mithramycin fluorescence in the clinical detection and quantitation of aneuploidy by flow cytometry.

1982 ◽  
Vol 30 (4) ◽  
pp. 317-322 ◽  
Author(s):  
R E Cunningham ◽  
K S Skramstad ◽  
A E Newburger ◽  
S E Shackney
Keyword(s):  
Flow Cytometry ◽  
Room Temperature ◽  
Human Melanoma ◽  
Time Dependent ◽  
Clinical Samples ◽  
Fixation Time ◽  
C Cells ◽  
Temperature Dependent ◽  
Clinical Detection

Ethanol-fixed cells stored at 4 degrees C exhibit fixation time-dependent hyperchromatism in comparison with freshly fixed cells when stained with mithramycin and examined by flow cytometry. This hyperchromatism has been found to be temperature-dependent, developing fully within 72 hr at room temperature, and within 2 hr at 37 degrees C. Cells from normal donors that are stained with mithramycin exhibit spurious aneuploid peaks. These spurious aneuploid peaks can be eliminated by incubating ethanol-fixed cells at 37 degrees C for 2 hr prior to staining; true aneuploidy is not affected by this procedure. In rare instances, cytoplasmic fluorescence can be observed in mithramycin-stained cells. In addition, unexplained hypochromatism and hyperchromatism can be observed in some clinical samples, particularly in human melanoma. The effects of these unexplained staining artifacts can be minimized or eliminated by adopting strict criteria for the clinical detection of aneuploidy by flow cytometry.

Quantification of fetomaternal hemorrhage by fluorescence microscopy is equivalent to flow cytometry

Transfusion ◽  
2002 ◽  
Vol 42 (7) ◽  
pp. 947-953 ◽  
Author(s):  
Nadin Ochsenbein‐Imhof ◽  
A.F. Ochsenbein ◽  
B. Seifert ◽  
Albert Huch ◽  
Renate Huch ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Fluorescence Microscopy ◽  
Fetomaternal Hemorrhage

Fetal cells in the maternal circulation: isolation by multiparameter flow cytometry and confirmation by polymerase chain reaction

1991 ◽  
Vol 6 (10) ◽  
pp. 1466-1469 ◽  
Author(s):  
Stephen Wachtel ◽  
Sherman Elias ◽  
James Price ◽  
Gwendolyn Wachtel ◽  
Owen Phillips ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Polymerase Chain Reaction ◽  
Multiparameter Flow Cytometry ◽  
Chain Reaction ◽  
Maternal Circulation ◽  
Fetal Cells ◽  
Polymerase Chain

scholarly journals Applications of Flow Cytometry to Clinical Microbiology

2000 ◽  
Vol 13 (2) ◽  
pp. 167-195 ◽  
Author(s):  
Alberto Álvarez-Barrientos ◽  
Javier Arroyo ◽  
Rafael Cantón ◽  
César Nombela ◽  
Miguel Sánchez-Pérez
Keyword(s):  
Flow Cytometry ◽  
Clinical Microbiology ◽  
Ex Vivo ◽  
Analytical Techniques ◽  
Clinical Samples ◽  
Clinical Microbiology Laboratory ◽  
Commercial Kits ◽  
Antimicrobial Treatments ◽  
Near Future

SUMMARY Classical microbiology techniques are relatively slow in comparison to other analytical techniques, in many cases due to the need to culture the microorganisms. Furthermore, classical approaches are difficult with unculturable microorganisms. More recently, the emergence of molecular biology techniques, particularly those on antibodies and nucleic acid probes combined with amplification techniques, has provided speediness and specificity to microbiological diagnosis. Flow cytometry (FCM) allows single- or multiple-microbe detection in clinical samples in an easy, reliable, and fast way. Microbes can be identified on the basis of their peculiar cytometric parameters or by means of certain fluorochromes that can be used either independently or bound to specific antibodies or oligonucleotides. FCM has permitted the development of quantitative procedures to assess antimicrobial susceptibility and drug cytotoxicity in a rapid, accurate, and highly reproducible way. Furthermore, this technique allows the monitoring of in vitro antimicrobial activity and of antimicrobial treatments ex vivo. The most outstanding contribution of FCM is the possibility of detecting the presence of heterogeneous populations with different responses to antimicrobial treatments. Despite these advantages, the application of FCM in clinical microbiology is not yet widespread, probably due to the lack of access to flow cytometers or the lack of knowledge about the potential of this technique. One of the goals of this review is to attempt to mitigate this latter circumstance. We are convinced that in the near future, the availability of commercial kits should increase the use of this technique in the clinical microbiology laboratory.

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